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31.
32.
Earlier reports suggested that the adenosine monophosphate (AMP)- and the p-nitrophenyl phosphate (pNPP)-hydrolyzing activities of Dictyostelium discoideum membrane preparations are due to different proteins. These results have been apparently contradicted by the recent purification to homogeneity of the two activities from culmination phase cells as a single protein [D. R. Armant and C. L. Rutherford (1981) J. Biol. Chem. 256, 12710-12718]. Results presented here from studies on the activities of vegetative cells support the concept of a single protein. Nondenaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of Triton X-100 extracts of cell membrane preparations of D. discoideum showed identical migration of pNPPase and AMPase activities. Furthermore, the previously reported different pH optima of the two activities was due to the fact that pH optima are dependent upon the substrate concentration, and the selective solubilization of AMPase from membrane preparations by phospholipase C can probably be accounted for by the finding that phospholipase C preparations from the same commercial source contain 5'-nucleotidase activity. Moreover, there are alterations in the Km and the stability of both AMPase and pNPPase in a strain with a mutationally altered alkaline phosphatase, further supporting the concept that the two activities are due to a single protein. Both substrates serve as transphosphorylation donors demonstrating that the enzyme activity is mechanistically an alkaline phosphatase. 相似文献
33.
Temporal Profiles of Proteins Responsive to Transient Ischemia 总被引:4,自引:3,他引:1
The responses of long and short half-lived proteins to ischemia were measured in rat brain during 6 days of recovery from 30 min of transient forebrain ischemia produced by four-vessel occlusion. At the end of the ischemic interval, the neocortical activities of four vulnerable enzymes [ornithine (ODC) and S-adenosylmethionine (SAMDC) decarboxylases, and RNA polymerases I and II] were unchanged, but within 30 min of reperfusion, their activities dropped by 25-50%. The loss of substance P in the striatum and substantia nigra was slower, reaching about 50% by 12 h. On the other hand, the activities of 5 long half-lived enzymes did not change in the neocortex at 5 and 15 h of reperfusion and regional protein concentrations were essentially unaffected over 6 days survival. The rate and extent of normalization of the amounts or activities of the vulnerable proteins varied. RNA polymerase II and ODC activities were restored within 4 h, and ODC showed a biphasic increase in activity, with peaks at 10 h and 2-3 days. RNA polymerase I and SAMDC activities were restored by 18 h and 5 days, respectively, whereas substance P concentrations did not completely recover, even at 6-15 days. The greater the regional reduction of blood flow during ischemia, the larger the net change (gain or loss) of SAMDC or ODC activity and the longer the time required to normalize the activities of these enzymes. The average rate of proteolysis, assessed by measuring the rate of clearance of 14C from protein prelabeled with [14C]bicarbonate, was abnormal during the first 2 days of reperfusion. Postischemic changes in both protein synthesis and degradation could affect the amounts of some of the proteins responsive to transient ischemia. 相似文献
34.
Deprenyl Protects Dopamine Neurons from the Neurotoxic Effect of 1-Methyl-4-Phenylpyridinium Ion 总被引:3,自引:2,他引:1
1-Methyl-4-phenylpyridinium ion (MPP+) is the product of the metabolic oxidation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) by monoamine oxidase (MAO). MPP+ is toxic to 3,4-dihydroxyphenylethylamine (dopamine, DA) neurons in explant cultures of rat embryonic midbrain. Addition of 2.5 microM MPP+ to the feeding medium for 6 days results in significant reduction of the DA levels in the cultures (to 19% of control) as well as in the uptake of [3H]DA (to 32% of control). When the cultures are treated with the MAO inhibitor deprenyl (10 microM) 24 h prior to and during exposure to MPP+, the DA neurons are protected from the toxicity of the drug. In the combined deprenyl plus MPP+ treatment, the levels of DA in the cultures remain at the control range and the [3H]DA uptake is reduced to only 73% of control. These results indicate that MAO is involved in the toxicity of MPP+ on DA neurons. 相似文献
35.
To identify sequences within the adenovirus-5 E3 promoter necessary for E1A trans-activation, a series of promoter deletion mutants were constructed and analysed. A region between positions -82 and -105 was shown to be critical both for E1A induced expression as well as uninduced expression. The importance of this region was confirmed by constructing hybrid promoters consisting of E3 and Herpes simplex virus thymidine kinase sequences. The E1A insensitive tk promoter could be converted to an E1A sensitive promoter by replacing sequences upstream of position -79 with the corresponding region of the E3 promoter. This critical region of the E3 promoter contains a sequence 5' AGATGACTA3' which is also present in important upstream regions of the E2A and E4 promoters. 相似文献
36.
Gerald S. Manning 《Cell biochemistry and biophysics》1985,7(3):177-184
The equilibrium trajectory of the axis of a rod subject to an externally imposed curved potential energy trough tends to conform
to the shape of the curved trough, but also tends to be straight because of elastic resistance to bending. The actual path
of the axis is a balance between the two extremes. We consider a potential energy trough centered along a circular arc of
radiusR. For a rod of small length compared toR, we show that the axis at equilibrium forms an arc of a circle of radius greater thanR. The value of the radius of the axial path depends on the relative values of the Hooke’s Law bending constant for the rod
and the depth and width of the trough. Motivation for the calculation is provided by nucleosomal DNA, which conforms to the
surface of a roughtly cylindrical histone core at physiological ionic strength, but is observed to unwind into a partially
extended conformation at very low ionic strength. We suggest that the rigidity to bending of short DNA segments becomes sufficiently
great at low ionic strength to overcome attractive interactions with the histone surface. Alternately, of course, if during
the cell cycle mutually attractive forces between DNA and histone core are weakened at constant ionic strength, the same type
of unfolding would be expected to occur as the strength of the DNA-histone contacts drops below the level required to overcome
elastic resistance to bending of the DNA rod. 相似文献
37.
The effects of mineral salts constituting water hardness on fecundity ofDaphnia magna were assessed. Of the salts tested, increased concentrations of NaHCO3 and MgSO4 had no effect on fecundity, CaSO4 significantly increased fecundity, and KCl significantly reduced fecundity. The number of offspring produced per daphnid
was correlative to the CaSO4 concentration at CaSO4 concentrations between 91 and 2100 mg/ℓ. The effects of CaSO4 on daphnid fecundity could influence the interpretive outcome of industrial wastewater toxicity tests using this species
when the waste and dilution waters contain different concentrations of CaSO4. It is recommended that when performing these tests, dilution water be sampled at the intake site of the industry's water
source, thus assuring initial comparability of the waste and dilution waters. The CaSO4 content of the water prior to and after industrial use should be determined to identify any alterations of CaSO4 concentration during use. Identification of CaSO4 concentration differences can aid in the interpretation of effects associated with the wastewater. 相似文献
38.
The rise time, of Signal IIf and the decay time of P-680+ have been measured kinetically as a function of pH by using EPR. The Photosystem II-enriched preparations which were used as samples were derived from spinach chloroplasts, and they evolved oxygen before Tris washing. The onset kinetics of Signal IIf are in agreement, within experimental error, with the fast component of the decay of an EPR signal attributable to P-680+. The signal IIf rise kinetics also show good agreement with published values of the pH dependence of the decay of P-680+ measured optically (Conjeaud, H. and Mathis, P. (1980) Biochim. Biophys. Acta 590, 353–359). These results are consistent with a model where the species Z (or D1) responsible for Signal IIf is the immediate electron donor to P-680+ in tris-washed Photosystem II fragments. 相似文献
39.
Gerald Rosen 《Bulletin of mathematical biology》1983,45(2):151-153
First-order spatial gradients are reliquished in the Schrödinger-Bloch equation for bacterial chemotaxis if and only if the flux coefficient-motility ratio equals 2, the precise value measured in recent experiments onEscherichia coli attracted by oxygen. Moreover, for δ/μ=2 the Schrödinger-Bloch function Ψ is simply equal to the number of bacteria cells per unit volume divided by the chemoattractant concentration. 相似文献
40.
Peter E. Prevelige Gerald D. Fasman 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,739(1):85-96
Core histones, (H2A,H2B,H3,H4)2, were reconstituted with the synthethic polynucleotides poly(dA-dT)·poly(dA-dT) and poly(dG-dC)·poly(dG-dC) to yield synthetic chromatins containing 200 basepairs per octamer. These synthetic chromatins displayed a 36% decrease in the circular dichroism (CD) peak ellipticity from the value of the polynucleotide free in solution; the poly(dA-dT)·poly(dA-dT)/chromatin showed an increase in the complexity of the thermal denaturation profile compared to that of the polynucleotide. Both the temperature of maximum for each transition (Tm) and the relative amount of poly(dA-dT)·poly(dA-dT) in the synthetic chromatin melting in each of the four thermal transitions is a function of the ionic strength over the 0–5 mM sodium phosphate range (0.25 mM EDTA, pH 7.0); a shift of material toward higher melting transitions was observed with increasing ionic strength. The CD peak ellipticity value for both synthetic chromatins was ionic strength-independent over the 0–5 mM sodium phosphate range. These results are in contrast to those observed with stripped chicken erythrocyte chromatin (Fulmer, A. and Fasman, G.D. (1979) Biopolymers 18, 2875–2891), where an ionic strength dependence was found. Differences in the CD spectra between poly(dA-dT)·poly(dA-dT)/chromatin, poly(dG-dC)·poly(dG-dC)/chromatin and stripped chicken erythrocyte chromatin suggest subtle differences in assembly. Finally, the temperature dependence of the CD spectra of poly(dA-dT)·poly(dA-dT)-containing synthetic chromatin, which is similar to that for the polynucleotide, suggests the core histone bound polynucleotide has a large degree of conformational flexibility allowing it to undergo the premelt transition. 相似文献